In silico study to predict promiscuous T cell and B cell-epitopes derived from the vaccine candidate antigens of Plasmodium vivax binding to MHC class-II alleles

Malaria is one of the major causes of health and disability globally, even after tremendous efforts to eradicate it. Till date no highly effective vaccine is available for its control. The primary reason for the low efficacy of vaccines is extensive polymorphism in potential vaccine candidate antigen genes and HLA polymorphisms in the human population. This problem can be resolved by developing a vaccine using promiscuous peptides to combine the number of HLA alleles. This study predicted T and B cell epitopes (promiscuous peptides) by targeting PPPK-DHPS and DHFR-TS proteins of Plasmodium vivax, using different in silico tools. Selected peptides were characterized as promiscuous peptides on the basis of their immunogenicity, antigenicity and hydrophobicity. Furthermore, to confirm their immunogenicity, these peptides were utilized for molecular modelling and docking analysis. For determining the requisite affinity with distinct HLA Class-I, and HLA Class-II alleles, only five peptides for DHFR-TS and 3 peptides for PPPK-DHPS were chosen as promiscuous peptides. The D1 peptide has the maximum binding energy with HLA alleles, according to HLA-peptide complex modelling and binding interaction analyses. These findings could lead to the development of epitope-based vaccinations with improved safety and efficacy. These epitopes could be major vaccine targets in P. vivax as they possess a higher number of promiscuous peptides. Also, the B cell epitopes possess maximum affinity towards different alleles as analyzed by docking scores. However, further investigation is warranted in vitro and in vivo.

Enzyme inhibition and antibacterial potential of 4-Hydroxycoumarin derivatives

The 4-Hydroxycoumarin derivatives are known to show a broad spectrum of pharmacological applications. In this paper we are reporting the synthesis of a new series of 4-Hydroxycoumarin derivatives synthesized through Knovenegal condensation; they were characterized by using UV-Vis, FT-IR, NMR spectroscopies. The synthesized compounds were evaluated for antibacterial activity against Staphylococcus aureus and Salmonella typhimurium strains. The compounds (2), (3) and (8) showed favorable antibacterial activity with zone of inhibitions 26.5± 0.84, 26.0 ± 0.56 and 26.0 ± 0.26 against Staphylococcus aureus (Gram-positive) respectively. However, the compounds (5) and (9) were found more active with 19.5 ± 0.59 and 19.5 ± 0.32 zone of inhibitions against Salmonella typhimurium (Gram-negative). Whereas, in urease inhibition assay, none of the synthesized derivatives showed significant anti-urease activity; although, in carbonic anhydrase-II inhibition assay, the compound (2) and (6) showed enzyme inhibition activity with IC50 values 263±0.3 and 456±0.1, respectively.

Isolation of dihydrobenzofuran derivatives from ethnomedicinal species Polygonum barbatum as anticancer compounds

BACKGROUND: Ethnomedicinally, the family Polygonaceae is famous for the management of cancer. Various species of this family have been reported with anticancer potentials. This study was designed to isolate anticancer compounds from ethnomedicinally important species Polygonum barbatum. METHODS: The column chromatography was used for the isolation of compounds from the solvent fraction of P. barbatum. The characterization of isolated compounds was performed by various spectroscopic techniques like UV, IR, mass spectrometry and 1D-2D NMR spectroscopy. Keeping in view the ethnomedicinal importance of the family, genus and species of P barbatum, the isolated compounds (1-3) were screened for anticancer potentials against oral cancer (CAL-27) and lungs cancer (NCI H460) cell lines using MTT assay. Active compound was further investigated for apoptosis by using morphological changes and flow cytometry analysis. In vivo anti-angiogenic study of the isolated compounds was also carried using chorioallantoic membrane assay. Docking studies were carried out to explore the mechanism of anticancer activity. RESULTS: Three dihydrobenzofuran derivatives (1-3) have been isolated from the ethyl acetate fraction of P. barbatum. The structures of isolated compounds were elucidated as methyl (2S,3S)-2-(3,4-dimethoxyphenyl)-4-((E)-3-ethoxy-3-oxoprop-1-en-1-yl)-7-methoxy-2,3-dihydrobenzo-furan-3-carboxylate (1), (E)-3-((2S,3S)-2-(3,4-dimethoxyphenyl)-7-methoxy-3-(methoxy carbonyl)-2,3-dihydrobenzofuran-4-yl)acrylic acid (2) and (2S,3 S)-4-((E)-2-carboxyvinyl)-2-(3,4-dimethoxyphenyl)-7-hydroxy-2,3-dihydrobenzofuran-3-carboxylic acid (3). The compound 1 was found to be more potent with IC50 of 48.52 ± 0.95 and 53.24 ± 1.49 against oral cancer cells as compared to standard drug (IC50 = 97.76 ± 3.44 µM). Both compound also inhibited lung cancer cells but at higher concentrations. Morphological and flow cytometry analysis further confirms that compound 1 induces apoptosis after 24 to 48 h treatment. In antiangiogenesis assay, compounds 1, 2 and 3 exhibited IC50 values of 8.2 ± 1.1,13.4 ± 1.1 and 57.7 ± 0.3 µM respectively. The docking studies revealed that the compounds under study have the potential to target the DNA and thymidylate synthase (TS). CONCLUSION: Based on its overwhelming potency against the tested cell lines and in angiogenesis assay, compound 1 can be further evaluated mechanistically and can be developed as anticancer drug candidate.

The effect of propofol when injected at different speeds for induction of general anesthesia: an observational study

Background:The hypotensive effect of propofolis attributable to a decrease in sympathetic activity,direct vasodilatation and myocardial depression. The aim of the study wasto assessthe effect of propofol when injected at different speeds for induction of general anesthesia on the following parameters:blood pressure,time of induction of anesthesia,dose of propofol used.Methods:The present study was conducted in post Graduate Department of Anesthesia and Surgery, Govt. Medical College,Srinagar for a period of two years and included 90 patients from July 2014 to June16,the study was prospective one.Results:In our study patients divided into three groups with 30 patients in each group.The mean age in group P400,P600,P800 wasstatistically insignificant p>0.843.The mean weight in group P400, P600, P800 was statistically insignificant p>0.885.The mean height in group P400,P600,P800 was statistically insignificant p>0.748.The mean induction time in P400 and in P600 was statistically significant.The mean systolic blood pressure, pre and post induction in P400,in P600 and in group P800 was statistically significant. The mean diastolic blood pressure, in pre and post induction in P400,P600,P800 was statistically insignificant with a p>0.05.The mean arterial pressure in pre and post induction in P400,P600,P800 was statistically significant (p<0.05).The mean heart rate in pre and post induction was statistically insignificant. The mean oxygen saturation (%) pre and post induction was statistically insignificant. Conclusions: We concluded that induction dose required for loss of consciousness increased with a faster rate of infusion while time for induction was shorter in P800 compared to P400 and P600, and the decrease in mean blood pressure was less after induction in P400.Propofol injection should be slow enough to prevent any hemodynamic deterioration in anesthesia induction

Sedative and muscle relaxant activities of diterpenoids from Phlomidoschema parviflorum

Abstract Phlomidoschema parviflorum (Benth.) Vved. (Basionym: Stachys parviflora Benth.) Lamiaceae, have significance medicinal importance as it is used in number of health disorders including diarrhea, fever, sore mouth and throat, internal bleeding, weaknesses of the liver and heart genital tumors, sclerosis of the spleen, inflammatory tumors and cancerous ulcers. The present contribution deals with the sedative and muscle relaxant like effects of diterpenoids trivially named stachysrosane and stachysrosane, isolated from the ethyl acetate soluble fraction of P. parviflorum. Both compounds (at 5, 10 and 15 mg/kg, i.p) were assessed for their in vivo sedative and muscle relaxant activity in open field and inclined plane test, respectively. The geometries of both compounds were optimized with density functional theory. The molecular docking of both compounds were performed with receptor gamma aminobutyric acid. Both compounds showed marked activity in a dose dependent manner. The docking studies showed that both compounds interact strongly with important residues in receptor gamma aminobutyric acid. The reported data demonstrate that both compounds exhibited significant sedative and muscle relaxant-like effects in animal models, which opens a door for novel therapeutic applications.

Molecular Detection of Mycobacterium Tuberculosis Complex from Sputum of Clinically Suspected Tuberculosis Cases

Background: Tuberculosis (TB) is a communicable disease and it  ranks second of all infectious agents due to co-infection with HIV . The causative agent of tuberculosis is a group of mycobacteria known as Mycobacterium tuberculosis complex. Mycobacterium tuberculosis complex consist of M. tuberculosis, M. bovis, M. africanum, M. microti, M. canetti. In PCR study, Most commonly sensitivity is higher in smear positive samples (95-100%) rather than smear negative specimens (46-63%). OBJECTIVE: To detection of Mycobacterium tuberculosis complex by Line Probe Assay. Methods: The study was done from non-interventional approaching study of 50 suspected patients of tuberculosis had visited the TB chest/ DOTS centre. Sputum sample collected early morning in a wide- mouth container from the patients having history of cough more than 2 weeks. Methodology used Z-N staining and for detection of MTB complex was done using MTBDR plus assay, multiplex PCR DNA strip assay (Hain Lifescience, Nehren, Gernamy) which is commercially available. M. tuberculosis secreted an important protein is MPT64, a 24-kDa protein. The major culture filtrate protein (24-kDa) is MPT64 encoded by the RD2 region genes and has been exposed to be an exact antigen that differentiates the M. tuberculosis complex from the mycobacteria other than tuberculosis (MOTT) Species. Results: In 50 samples, out of which 10 (20%) were smear positive & 40(80%) were smear negative. Out of 10 smear positive, 9 (90%) were MTB (Mycobacterium tuberculosis) & 01 (10%) was NTM (Non-tuberculous Mycobacteria) by PCR method. Out of 40 smear negative, 30 (75%) were positive by PCR method. Out of 30, 28(93%) were MTB & 02(7%) were NTM. Rests of the 10(25%) samples were found negative for M. tuberculosis complex. Conclusions: This study proved that PCR is a specific and sensitive method in comparison of sputum microscopy after staining with Z-N technique and it helpful  the clinicians ability to diagnose and treat the patients on time. This will ensure early treat to patients and prevent further transmission of disease.

Bacteriological Profile and Antibiogram of Gram Negative Bacteria Isolated from Blood Culture

Introduction: Blood stream infections are one of the important cause of morbidity and mortality all over the world. Bacteraemia ranges from self-limiting infections to life-threatening septicaemia that requires rapid and aggressive antimicrobial treatment. The mortality rate ranges from 20% to 50% in cases of bacteraemia infections. Aim and Objective: The present study was undertaken to know the profile of gram negative organism causing  bacteraemia with their Antibiogram from suspected cases. Material and Method: During a one-year period, 400 blood samples were taken from bacteraemia suspected patients. Blood culture was done by using BacT/Alert 3D system.  Further identification of organism was done by different biochemical test. Antimicrobial sensitivity pattern was determined by Kirby Bauer Disc Diffusion method according to CLSI guidelines. Result: Out of 400 samples, the total number of culture positive cases were found to be 131 giving culture positive rate of 32.75%. Gram positive organism were more than gram negative organism, constituting about 75 (57.69%) of total isolates. 56(42.74%) Gram negative organism were isolated in this study. Most frequent pathogen identified among gram negative bacteria were Klebseilla 24(42.8%), followed by E. coli 18(32.14%), Acinetobacter 10(17.85%), Pseudomonas 2(3.57%) and Salmonella 2(3.57%) respectively. Isolated gram negative organism was highly sensitive to Polymyxin B 51(91.07%). After Polymyxin B isolated gram negative bacteria show high sensitivity for Levofloxacin(60.71%), Cefixime (57.78%), Gentamicin, Meropenem, Piperacillin/tazobactum (50%), Cefepime (44.64%) with least sensitivity for Ampicillin/Sulbactum (14.28%). Conclusion: The present study provides information about gram negative pathogens responsible for blood    stream infection along with their sensitivity towards commonly used antimicrobial. Antibiotic sensitivity pattern of isolates provides useful guidelines to clinicians in initiating empiric therapy and help in management of blood stream infections.

Prevalence of Urinary Tract Infection in Patients attending Teerthanker Mahaveer Medical College & Research Centre and Hospital Moradabad

Background: It has been estimated that symptomatic urinary tract infections (UTI) occurs in around 7 million patients visiting to emergency units and 100,000 hospitalizations annually. It continues to be the most common cause of infection in hospitalized patients accounting approximately 40% of hospital acquired infections, and is the second most common cause of bacteraemia in hospitalized patients. Objective: a) To study the prevalence of UTI and its etiology in patients coming to the hospital. b) To study the susceptibility pattern of the isolate. Methods: The study was performed on 630 midstream urine positive samples taken from all age group patients, either male or female. Samples were collected prior administration of antibiotics in a sterile container. In case of cathetized patients, it is collected directly from catheter. Samples were inoculated on CLED agar. By colony count the stage of bacteraemia was assessed. To know the causative organism , colony morphology as well biochemical test were done. Results: The prevalence of Urinary Tract Infections (UTI) was evaluated in 630 patients attending Teerthanker Mahaveer Medical College & Research Centre, Moradabad for the duration of one year from February 2016 to January 2017. Results showed 215 (34.12%) patients were positive. Out of 215 positive cases 41(19.06%) were gram positive organisms and rest 174 (80.94%) were gram negative organisms. The most common pathogenic organisms were Escherichia coli accounting for 98(45.19%) urinary isolates. Among gram positive organism Enterococcus 26(12.09%) were the most common. In-vitro antibiotic susceptibility tests revealed that the gram negatives bacteria were sensitive to quinolones and Carbapenems, while the gram positive isolates were sensitive to glycopeptides antibiotics. Conclusions: The findings suggested the need for regular screening for the presence of symptomatic or asymptomatic bacteriuria for different populations and constant monitoring of susceptibility to commonly used anti-microbial agents.

In Vitro Assessment of Antileishmanial Activity of Natamycin and Nystatin

ABSTRACT The present study was aimed to evaluate the in vitro antileishmanial activity of four different concentrations of natamycin and nystatin by using MTT 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide reduction assay. In vitro antileishmanial activity revealed that the IC50 of natamycin (80.49 μg/ml) and nystatin (105.7 μg/ml) was less than that of sodium stibogluconate (127.9 μg/ml), and more than amphotericin B (18.91 μg/ml).

Isolation, Identification and Drug Resistance Testing of Mycobacterium tuberculosis by Recent Diagnostic Modalities at Teaching Hospital in Moradabad (UP).

Objective: Tuberculosis is the second leading cause of death in developing countries among all infectious diseases. Globally, drug resistance strain of Mycobacterium tuberculosis is a public threat. Due to diagnostic delay, inadequate infection control and poor drug supply there is a emergence of MDR- TB and XDR- TB. Our aim was to isolate and identify the drug resistant strain of M. tuberculosis by using newer diagnostic modalities. Methods: Sputum sample and BAL fluid from 70 suspected cases were collected and analysed for Mycobacterium by Ziehl – Neelsen staining and liquid culture with molecular detection of drug resistant strain of M. tuberculosis. Result: In our study, among the 70 patients 27 (38.5%) were positive for AFB by microscopy. On testing for Mycobacterium by BacT/Alert 3D system, 54 were found to be positive. On performing further identification and susceptibility of 54 isolates towards rifampicin and isoniazid by molecular method, 5 isolates (9.25%) were resistant to both rifampicin and isoniazid confirming as multidrug resistant. 5 isolates (9.25%) were sensitive to rifampicin and resistant to isoniazid and 2 isolates (3.70%) were resistant to rifampicin and sensitive to isoniazid. Whereas 5 isolates (9.25%) found to be negative for M. tuberculosis. Conclusion: Our investigation highlights the importance of newer diagnostic modalities for isolation and identification of drug resistant strain of M. tuberculosis. Which ensure early and accurate diagnosis of patients with prevention of further transmission of disease.

To study the Prevalence and Antibiotic Sensitivity Pattern of Isolated Bacteria from the Blood, Urine, CSF and Pus Samples in a Tertiary Care Teaching Hospital

Introduction: The worldwide escalation in both community- and hospital-acquired antimicrobial-resistant bacteria is threatening the ability to effectively treat patients, emphasizing the need for continued surveillance, more appropriate antimicrobial prescription, prudent infection control, and new treatment alternatives. Objective: To study the prevalence of bacteria from the different samples (Blood, Urine, CSF, PUS) and to examine the antibiotic sensitivity pattern of isolated organisms. Methods: Around n=150 samples of Urine, Blood, CSF and Pus sample were collected from the patient attending Teerthanker Mahaveer medical Hospital college and Research Centre. Results: Out of 150 clinical samples, highest number of isolates were gram-positive, Staphylococcus aureus n=47 (31.33%) followed by E. Coli n=37(24.66%), Klebseilla n=33(22.00%), Pseudomonas n=11(07.33%). S. aureus was highly sensitive to Gentamycin (88.09%) and least sensitive to Co-trimoxazole (14.28%). Tobramycin & Linezolid were 95.23% sensitive followed by Amikacin (90.47%), Meropenem (90.47%), Levofloxacin (88.09%). Conclusion: In our study Staphylococcus aureus to be most common isolates followed by Escherichia coli, Klebseilla, Pseudomonas, Citrobacter, Proteus.

Molecular Detection of Mycobacterium Tuberculosis Complex from Clinical Sputum Samples in Patients Attending Tertiary Care Centre in Uttar Pradesh Province of India.

Background: In the world, tuberculosis ranks second after HIV of all infectious agents leading cause of mortality and morbidity due to bacterial infections with HIV taking the first spot. India holds the global burden of TB in one fifth with more than 350,000 deaths each year. Though pulmonary TB (PTB) cases, account for the vast majority of the total TB burden, almost 10-15 per cent of total cases are extra-pulmonary infection. Methods: Mycobacterium Tuberculosis complex were detected in 50 clinical sputum samples by using Polymerase chain Reaction (PCR) and BacT/Alert. Results: In our study, 50 sputum clinical samples were taken, out of which 11(22%) were smear positive & 39 (78%) were smear negative. Out of 11 smear positive 10 (90%) were MTB (Mycobacterium Tuberculosis) & 01 (10%) was NTM (Non-Tuberculous mycobacteria) and in 39 smear negative,15 (38.47%) were M. tuberculosis & 02 (5.12%) were NTM and 22 (56.41%) samples were negative by using PCR. By BacT/Alert 3D system, out of 50 clinical samples only 15(30%) samples were positive and 35 (70%) samples were negative for M. tuberculosis complex. Conclusions: It is concluded that result obtained from our study, Mycobacterium tuberculosis complex was detected by PCR from clinical samples has high specificity (99%) and sensitivity (95%) than BacT/Alert 3D system.

Isolation and Identification of Organism from Throat Swab Along with Sensitivity Pattern from the Patients in North India.

Background: Sore throat is the one of the commonest complaint of the patients in ENT OPDs. The prevalent cause of sore throat is in India is group A streptococci (GAS).But physicians across the country underestimate the real cause of the sore throat and prescribe the irrational use of antibiotic leading to development of resistance towards the antibiotics by the pathogens. Methods: The100 throat swab samples were collected with all aseptic precautions from sore throat patients in ENT OPD and also from other department of microbiology from March 2014 to December 2015. And were sent to microbiology department for throat swab culture & Sensitivity reporting. Result: A total of 100 patients suffering from sore throats were included in this study of which 57 were males and 43 were females. Out of 100 patients 35 were pathogenic 59 were nonpathogenic & there were no growth in 6 patients. The age range of study is from 2 years to 70 years. The isolated organism were E.Coli, Klebsiella pneumoniae, Streptococcus pneumoniae, Staph aureus,and alpha haemolytic streptococci were found out of 35pathogenic organism. Biomodal peak of more pathogenic growth was observed in the month of September, November and December. Culture sensitivity reports showed high sensitivity in of various pathogens towards erythromycin (mostly), Prisinomycin ,Cotrimoxazole, Linezolid,Vancomycin ,Cefaperazone, Polymyxin B Norfloxacin, Ampicillin and resistance towards Chloramphenicol,Gentamycin, Ampicillin, Cefazolin, Amoxicillin etc. Conclusion: This study gives us an insight to the current state of causative pathogens and their antimicrobial sensitivity from throat swab in Teerthanker Mahaveer Hospital. Alpha haemolytic Streptococci and staphylococcus aureus were the commonest organism isolated from throat swab.

Fungi as chemical industries and genetic engineering for the production of biologically active secondary metabolites

Fungi is somewhere in between the micro and macro organisms which is a good source of producing biologically active secondary metabolites. Fungi have been used as tool for producing different types of secondary metabolites by providing different nutrients at different laboratory conditions. The fungi have been engineered for the desired secondary metabolites by using different laboratory techniques, for example, homologous and heterologous expressions. This review reported how the fungi are used as chemical industry for the production of secondary metabolites and how they are engineered in laboratory for the production of desirable metabolites;also the biosynthetic pathways of the bio-organic-molecules were reported.

Polymorphism in merozoite surface protein-1 gene in north & northwest indian field isolates of Plasmodium vivax.

Background & objective: Merozoite surface protein-1 of Plasmodium vivax (Pvmsp-1) is a strong vaccine candidate against asexual blood stages. Extensive polymorphism in msp-1 gene has been reported in P. vivax isolates from different geographical regions which is necessary before a field trial of any malaria vaccine based on msp-1 is undertaken. There are only a few reports available on polymorphism in msp-1 gene in Indian field isolates of P. vivax. The aim of the present study was therefore to investigate the polymorphism in Pvmsp-1 gene in 25 isolates of P. vivax collected from malaria patients from regions of north and northwest India. Methods: Parasite DNA was extracted from whole blood samples collected in citrated anticoagulant. The polymorphic region-5, the most variable region of the Pvmsp-1 gene was amplified by PCR. The PCR products were further analyzed by restriction fragment length polymorphism (RFLP) using Mva-1 restriction enzyme. The DNA fragments obtained on PCR and RFLP were analyzed by agarose gel electrophoresis. Results: On the basis of PCR, significant size polymorphism was seen and 4 allelic types were observed amongst the 25 isolates. Further analysis by RFLP discriminated these 4 allelic types into 9 sub-allelic types indicating that PCR-RFLP can be a good tool to study polymorphism in msp-1 gene of Plasmodium. Interpretation & conclusion: Marked genetic polymorphism was observed in msp-1 gene among the isolates of P. vivax. These observations stress the need to study larger numbers of isolates from different regions of India. The findings could have important implications on the vaccine development strategies for P. vivax.

Genetic Polymorphism in msp-2, ama-1 and csp genes in Plasmodium falciparum field isolates from north and north-western India.

Background & objectives: Malaria is a major public health problem in tropical and sub-tropical countries. Malaria vaccine is highly desirable as an adjunct to existing malaria control measures. The polymorphism in vaccine candidate antigens might be a hurdle in developing an effective vaccine. Merozoite surface protein-2, apical membrane antigen-1 and circumsporozoite protein of Plasmodium falciparum are vaccine candidate antigens. The aim of this study was to detect extent of genetic polymorphism in potential vaccine candidate antigen genes, i.e. msp-2, ama-1 and csp of P. falciparum isolates prevalent in northern and north-western parts of India. Methods: Overall 88 parasite isolates of P. falciparum were collected during July 1998–March 2002 from different parts of northern and north-western India. DNA was extracted and analyzed for genetic polymorphism by PCR-RFLP method. For msp-2 gene, family-specific (FC-27 and 3D7) nested PCR was also performed. Results: PCR showed size polymorphism in all the target genes. Three alleles were observed in msp-2 and ama-1, while only two in csp. RFLP of ama-1 and csp with Dra-1 and Ssp-1 endonucleases respectively, failed to differentiate isolates in sub-allelic types, while Hinf-I digestion of msp-2 amplicons differentiated three alleles into two distinct allelic families, i.e. FC-27 and 3D7. The allelic family-specific PCR generally confirmed the results of PCR-RFLP except in a few isolates, which showed mixed (two) clones of msp-2 gene. Interpretation & conclusion: There was extensive polymorphism in msp-2 gene while ama-1 and csp genes showed low polymorphism which may be due to the functional constraints of these proteins. The low level transmission of malaria in the study area may also be a factor for low polymorphism.

Genetic polymorphism in Plasmodium falciparum vaccine candidate antigens.

Malaria is still a major public health problem in many tropical and subtropical countries. Malaria vaccine is highly desirable as an adjunct to existing malaria control measures. The polymorphisms in malaria vaccine candidates antigens might be a hurdle in developing an effective vaccine. The present article reviews the genetic polymorphism in several antigens expressed on the parasite surface, which are targets for immunological responses of the host and are good candidates for vaccine development against P. falciparum. Variable regions of most genes are generally dimorphic probably as a result of intragenic recombinations. Each allele in turn shows polymorphism resulting from point mutations, or other mechanisms. Several antigens like merozoite surface protein-1 and 2 (MSP-1 and MSP-2) and S antigen show high polymorphism while in others like circumsporozoite protein (CSP), apical membrane antigen-1 (AMA-1) and erythrocyte binding antigen-175 (EBA-175) functional constraints limit the degree of polymorphism. Polymorphism reported in these genes is discussed.

Drug resistance in malaria.

Ever since the discovery of the first case of chloroquine resistance along the Thai-Combodian border in the late 1950s, Southeast Asia has played an important role as a focus for the development of drug resistance in Plasmodium falciparum. Although the first case of quinine resistance had been reported much earlier from South America, the onset of chloroquine resistance marked the beginning of a new chapter in the history of malaria in Southeast Asia and by 1973 chloroquine finally had to be replaced by the combination of sulphadoxine and pyrimethamine (SP) as first line drug for the treatment of uncomplicated malaria in Thailand and more than 10 African countries have also switched their first line drug to SP. In 1985, eventually SP was replaced by mefloquine. The rapid development of resistance to this new drug leads to the introduction of artemisinin as a combination drug in the mid-1990s. It is mandatory to mention here that therapeutic regimens for prevention and treatment of chloroquine-resistant P. falciparum are associated with higher costs and side-effects compared to chloroquine. Additionally, some of these alternative treatments are associated with more side-effects, take longer time for cure and are more difficult to comply with than chloroquine. Urgent efforts are needed to identify effective, affordable, alternative antimalarial regimens. Molecular markers for antimalarial resistance have been identified, including pfmdr-1 and pfcrt polymorphisms associated with chloroquine resistance and dhfr and dhps polymorphisms associated with SP resistance. Polymorphisms in pfmdr-1 may also be associated with resistance to chloroquine, mefloquine and artemisinin. Use of such genetic information for the early detection of resistance foci and future monitoring of drug resistant malaria is a potentially useful epidemiological tool, in conjunction with the conventional in vitro and in vivo drug sensitivity assessments. The purpose of this review is to describe the state of knowledge regarding drug resistant malaria and to outline the changing patterns of drug resistance including its determinants, current status in diverse geographical areas, molecular markers and their implications to limit the advent, spread and intensification of drug resistant malaria.

Seroprevalence of rubella in pregnant women in Kashmir.

The present study was conducted to determine the seroprevalence of rubella in pregnant women in Kashmir. One thousand nine hundred and eighteen pregnant women in age group of 20-35 were divided into two groups. Group I (n=892) consisted of women with previous history of repeated abortion (507), single abortion (220), intrauterine death (149), stillbirth (14) and premature delivery (2). Group II (n=1028) pregnant women with previous normal delivery. A total of 1918 pregnant women were screened for rubella IgM antibodies out of which 16.74% were positive. In women with bad obstetric history (Group I) 26.12% were positive as compared to 8.96% in women with no significant obstetric history (Group II). The IgM antibody positivity was higher in women with previous history of intrauterine death (IUD) 58.38% followed by stillbirth 57.14%, premature delivery 50%, abortion 21.8% and recurrent abortion 17.55%. The high prevalence of disease in this region demands urgent needs for prevention. Moreover antenatal cases should be screened as early diagnosis and time intervention will help in proper management of these cases.